Miller Sean J, Rothstein Jeffrey D
Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, United States of America.
The Brain Science Institute at Johns Hopkins University, Baltimore, MD, United States of America.
PLoS One. 2016 Aug 5;11(8):e0160391. doi: 10.1371/journal.pone.0160391. eCollection 2016.
We utilized the recently published method of passive CLARITY to explore brain astrocytes for the first time with our optimized method. Astrocytes are the fundamental cells in the brain that act to maintain the synaptic activity of neurons, support metabolism of all neurons, and communicate through extensive networks throughout the CNS. They are the defining cell that differentiates lower organisms from humans. From a disease vantage point they are the principal cause of brain tumors and the propagator of neurodegenerative diseases like amyotrophic lateral sclerosis. New methods to study these cells is paramount. Our modified use of CLARITY provides a new way to study these brain cells. To reduce cost, speed up tissue clearing process, reduce human handling error, and to retrieve quantifiable data from single confocal and pseudo-super resolution microscopy we modified and optimized the original protocol.
我们首次运用最近发表的被动CLARITY方法以及我们优化后的方法来探索脑星形胶质细胞。星形胶质细胞是大脑中的基础细胞,其作用是维持神经元的突触活动、支持所有神经元的新陈代谢,并通过广泛的网络在整个中枢神经系统中进行通信。它们是区分低等生物和人类的标志性细胞。从疾病的角度来看,它们是脑肿瘤的主要成因以及诸如肌萎缩侧索硬化等神经退行性疾病的传播者。研究这些细胞的新方法至关重要。我们对CLARITY的改进应用为研究这些脑细胞提供了一种新途径。为了降低成本、加快组织透明化过程、减少人为操作误差,并从单共聚焦和伪超分辨率显微镜中获取可量化的数据,我们对原始方案进行了修改和优化。